Bio-Rad Bio-Plex Pro™ Magnetic Cell Signaling Assays - Manual

This guide describes how to prepare and run a full 1 x 96-well flat bottom
plate using a magnetic assay workflow. For more information on a given
step, refer to the corresponding section of the complete instruction manual.
New users can download the manual, which includes detailed instructions
and a list of kit components, at www.bio-rad.com/bio-plex.
Sample Preparation
1. Prepare lysates from cell culture or tissue samples. To remove insoluble
debris from lysates (≤2 ml), centrifuge at 15,000 x g for 10 min at 4˚C in
a benchtop microfuge. (Section 1 of the complete instruction manual)
2. Measure protein concentration using the Bio-Rad ® DC ™ protein assay.
3. Adjust protein concentration to 20–200 µg/ml in Bio-Plex ® cell lysis buffer
containing cell lysis factor QG and PMSF. For samples with unknown
target expression or phosphorylation levels, start with 200 μg/ml protein.
4. If necessary, store lysates at –70˚C.
5. Thaw frozen lysates and keep on ice. Centrifuge again as above and
bring to room temperature (RT) immediately before use.
Bio-Plex Pro ™ Assays
Quick Guide 3
IMPORTANT! Pay close attention to vortexing, shaking, and incubation instructions.
Deviation from the protocol may result in low assay signal and assay variability. Do Not
mix phosphoprotein assays with corresponding total target assays (e.g., phospho-Akt
and total Akt). Use only the reagents supplied with cell signaling assay kits.
For Use With:
Product Family
Instruction Manual #
Bio-Plex Pro cell signaling assays 10024929